CN103695337B - Adopt the method for thermophilic microorganism microbial inoculum process antibiotic bacterium slag - Google Patents
Adopt the method for thermophilic microorganism microbial inoculum process antibiotic bacterium slag Download PDFInfo
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Abstract
The invention provides a kind of method adopting thermophilic microorganism microbial inoculum process antibiotic bacterium slag, in antibiotic bacterium slag, add thermophilic microorganism microbial inoculum, make the minimizing of antibiotic bacterium slag, and make the residual reduction of the antibiotic in bacterium slag.Under aerobic fermentation condition, antibiotic bacterium slag decrement more than 85%, the antibiotic residue removal rate in bacterium slag reaches more than 90%.The advantage of present method is that antibiotic bacterium slag minimizing degree is high, and residue removal rate is high.The present invention is by antibiotic bacterium slag decrement, and the bacterium slag after decrement burns again, has greatly saved burning cost.This method is simple, and easy handling, floor space is little, and cost of investment is low, can not produce secondary pollution to environment, has obvious economic benefit, social benefit and environmental benefit.
Description
Technical field
The invention belongs to biological degradation field, specifically, relate to a kind of method adopting thermophilic microorganism microbial inoculum process antibiotic bacterium slag.
Background technology
Antibiotic plays huge effect in the survival and development and Animal husbandry production of the mankind.Up to the present, antibiotic its Application Areas still cannot replace by other materials.Antibiotic bacterium slag is the inevitable outcome of antibiotic production process.These bacterium slags, because of containing rich in protein, polysaccharide, amino acid, after being used the technical finesses such as dry processing or as feed, fodder additives, or are fully utilized as the raw material of fertilizer, production composite fertilizer before 2008 always.But because antibiotic is residual higher in antibiotic bacterium slag, as the residual about 2 ~ 2.5mg/g of cephalosporin in bacterium slag, the residual about 1 ~ 2mg/g of penicillin in bacterium slag, the residual about 0.2 ~ 0.9mg/g of erythromycin in bacterium slag, the residual about 0.5 ~ 2.1mg/g of terramycin in bacterium slag, the residual about 0.5 ~ 2mg/g of tsiklomitsin in bacterium slag.These residues finally all can be entered in animal body with in human body by food chain.The food that long-term absorption contains amino glucosides antibiotics residual can cause renal toxicity and ototoxicity; The food that long-term absorption contains cephalosporin penicillins antibiotic residual causes asthma, purple plague purpura; It can make pathogenic bacteria resistance to drugs increase in addition, thus brings prevention and the difficulty for the treatment of some human diseases and livestock poultry disease, finally causes pasting medical help; And they are made the numerous imbalances that Fertilizer application can cause the microorganism ecological environment in soil, soil fertility declines, and accelerates stony desertification Desertification; Affect the export trade of China's livestock product simultaneously.
After 2008, antibiotic bacterium slag is classified as Hazardous wastes by country, can only burn by dangerous waste disposal.China has become the maximum antibiotic production of raw medicine state in the whole world and export State, and bulk drug output has accounted for 80% of global ultimate production, and its middle outlet accounts for more than 70% of bulk drug market, the whole world.Within 2012, the whole nation produces about 140,000 tons, antibiotic, and its bacterium slag is more than 1,500,000 tons.The burning cost of bacterium slag has become the bottleneck of antibiotic industry survival and development.
To the process of antibiotic bacterium slag, all seeking more appropriate treatment approach always.Many scientific research institutions and pharmacy corporation have carried out the research of the disposal of antibiotic bacterium dregs innocent and different purposes.Environmental protection department also pays much attention to and encourages the exploitation of the new technologies such as antibiotic bacterium slag disposal.Such as, more successfully 1 is had) dregs of a decoction charing disposal; 2) biomass energy cogeneration technology.
Summary of the invention
The object of this invention is to provide a kind of method adopting thermophilic microorganism microbial inoculum process antibiotic bacterium slag.
In order to realize the object of the invention, first the present invention provides a kind of thermophilic microorganism composite fungus agent for the treatment of antibiotic bacterium slag, in described composite fungus agent, effective constituent comprises thermophilic ground bud bacillus (Geobacillussp.) UTM01 that deposit number is CGMCCNo.5641, deposit number is thermophilic ground bud bacillus (Geobacillussp.) UTM02 of CGMCCNo.5642, deposit number is genus bacillus (Bacillussp.) UTM03 of CGMCCNo.5643, deposit number is methylotrophic genus bacillus (Bacillusmethylotrophicus) UTM401 of CGMCCNo.5927, deposit number is anaerobic spore-bearing bacilli (Anoxybacillusmongoliensis) UTM501 of CGMCCNo.5928, deposit number is thermophilic double oxygen genus bacillus (Anoxybacilluspushchinoensis) UTM601 of CGMCCNo.5929, they are all the microorganisms filtered out from hot environment.
The present invention also provides the preparation method of described thermophilic microorganism composite fungus agent, comprises the step of the slant culture of bacterial strain, seed culture and fermentation culture, treats bacterium liquid OD
600>=1.9, merge each bacterium liquid, obtain thermophilic microorganism composite fungus agent.Viable count>=1 × 10 in described thermophilic microorganism composite fungus agent
11cFU/mL.
Slant medium: peptone 10g, yeast extract paste 10g, sodium-chlor 10g, agar 16 ~ 20g, water 1000mL, pH nature.
Seed culture medium: peptone 10g, yeast extract paste 10g, sodium-chlor 10g, water 1000mL, pH6.5 ~ 7.2.
Fermention medium: glucose 5g, yeast extract paste 10g, peptone 10g, sodium-chlor 10g, water 1000mL, pH6.0 ~ 7.0.
Shake-flask seed is cultivated: scrape lawn from each bacterial strain inclined-plane respectively and be inoculated in seed culture medium, cultivates 20 ~ 24 hours for 40 DEG C, shaking speed 200 ~ 220rpm.Treat each bacterium liquid OD
600>=1.5, and bottle.And the seed liquor after bottle is inoculated in fermentor tank.
Fermentor cultivation: the in-built 30L fermention medium of 50L fermentor tank, inoculum size 0.1 ~ 0.5%(v/v), cultivate 20 ~ 24 hours for 40 DEG C, fermentor tank rotating speed 100 ~ 200rpm, air flow 1:0.1 ~ 0.3, OD
600>=1.9 stop cultivating.
Microbial inoculum finished product: fermented liquid direct packaging 25kg/ bucket or packing 10kg/ bag after granulating.
The present invention also provides the method adopting thermophilic microorganism microbial inoculum process antibiotic bacterium slag, namely utilizes thermophilic microorganism composite fungus agent process antibiotic bacterium slag of the present invention.Antibiotic bacterium slag after process is by minimizing more than 85%, and the clearance that in antibiotic bacterium slag, antibiotic is residual can reach more than 90%.
Wherein, the moisture content of described antibiotic bacterium slag is adjusted to 55 ~ 65%, adds microbial inoculum 1L or 1kg in bacterium slag per ton.Described antibiotic bacterium slag includes but not limited to cephalosporin dreg, terramycin strain slag, erythromycin bacterium slag and lovastatin bacterium slag etc.
It utilizes described thermophilic microorganism composite fungus agent by aerobic static fermentation technical finesse antibiotic bacterium slag.Leavening temperature can reach more than 90 DEG C, and fermentation period is 15 ~ 20 days.Air flow is kept at 1:0.05 ~ 0.2(v/v) in fermenting process.
The method of employing thermophilic microorganism microbial inoculum process antibiotic bacterium slag provided by the invention, also comprises the step that the bacterium slag after to microbial inoculum process burns.
Particularly, the aerobic static fermentation technology of antibiotic bacterium slag of the present invention, the fermenter size of fermentation plant and number, depending on bacterium Slag treatment amount, are provided with and mix confounding, runway (Fig. 1,2).Each fermenter ground is provided with 1 ~ 3 chase, is covered with ventpipe in chase, ventpipe has many ventilating pits, uses blower fan air feed.Mixing confounding with forklift bacterium slag, moisture adjuster mixed be mixed even, make its moisture content in 55 ~ 65% scopes.1L or 1kg thermophilic microorganism composite fungus agent is added again, mixing by 1 ton of bacterium slag.The above-mentioned material forklift mixed moves on in fermenter, and placing height must not lower than 2.2M.Blower fan is adopted to be interrupted or uninterrupted air feed heap body.As 100 tons of heap bodies use power of fan to be about 4KW, full blast pressure 14kpa, air quantity 700M
3/ h.
Fermentation certain hour after adopt forklift with fall groove form to material turn over throw or adopt turnover throwing machine throwing is turned over to material.Turn over and throw number of times depending on fermentation situation, fermentation period about 3 ~ 4 times.
The change of temperature, water content and bacterium slag decrement and the residual relation of antibiotic in fermenting process:
Containing materials such as a large amount of protein, polysaccharide, amino acid and a small amount of antibiotic in bacterium slag.Microorganism utilizes these nutritive substances and other organism prolific heat of simultaneously releasing that heap temperature is raised gradually, and water content reduces gradually, and the bacterium quantity of slag reduces, the minimizing that antibiotic is residual.Ferment 3rd ~ 4 days heap temperatures 50 ~ 60 DEG C, moisture content about 45 ~ 52%, decrement 5 ~ 10%, clearance about 1 ~ 2%; Within 5th ~ 6 days, heap temperature rises 80 ~ 90 DEG C, and moisture contains about 48%, decrement 15 ~ 20%, clearance about 5%, within 7th ~ 8 days, temperature rises to 100 ~ 105 DEG C, and moisture is containing about 45%, decrement 40 ~ 50%, clearance about 20 ~ 30%, maintains 100 ~ 105 DEG C of high temperature for 9th ~ 16 days always, until moisture content is down to 30%, heap temperature starts to decline, fermentation ends, now bacterium slag decrement more than 85%, antibiotic clearance more than 90%, fermentation period 15 ~ 20 days (Fig. 3,4).
The invention provides one and add thermophilic microorganism microbial inoculum in antibiotic bacterium slag, make the minimizing of antibiotic bacterium slag, and make the antibiotic in bacterium slag remain the method for reduction.The present invention adopts the composite fungus agent of thermophilic microorganism as antibiotic bacterium Slag treatment bacterial strain, and it makes antibiotic bacterium slag decrement more than 85% under the condition of aerobic fermentation, and the antibiotic residue removal rate in bacterium slag reaches more than 90%.The advantage of present method is that antibiotic bacterium slag minimizing degree is high, and residue removal rate is high.The present invention is by antibiotic bacterium slag decrement, and the bacterium slag after decrement burns again, has greatly saved burning cost.This method is simple, and easy handling, floor space is little, and cost of investment is low, and can not produce secondary pollution to environment, has obvious economic benefit, social benefit and environmental benefit.
Accompanying drawing explanation
Figure 1 shows that the static fermentation groove that process antibiotic bacterium slag of the present invention uses.
Figure 2 shows that the ground air feed chase of fermenter of the present invention.
Fig. 3 and state when Figure 4 shows that antibiotic bacterium slag of the present invention ferments.
Embodiment
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.If do not specialize, the conventional means that technique means used in embodiment is well known to those skilled in the art, is raw materials usedly commercial goods.
Embodiment 1 is for the treatment of the thermophilic microorganism composite fungus agent of antibiotic bacterium slag and preparation thereof
In described composite fungus agent, effective constituent comprises thermophilic ground bud bacillus (Geobacillussp.) UTM01 that deposit number is CGMCCNo.5641, deposit number is thermophilic ground bud bacillus (Geobacillussp.) UTM02 of CGMCCNo.5642, deposit number is genus bacillus (Bacillussp.) UTM03 of CGMCCNo.5643, deposit number is methylotrophic genus bacillus (Bacillusmethylotrophicus) UTM401 of CGMCCNo.5927, thermophilic double oxygen genus bacillus (Anoxybacilluspushchinoensis) UTM601 of deposit number to be anaerobic spore-bearing bacilli (Anoxybacillusmongoliensis) UTM501 of CGMCCNo.5928 and deposit number be CGMCCNo.5929, they are all the microorganisms filtered out from hot environment.
Viable count>=1 × 10 in described thermophilic microorganism composite fungus agent
11cFU/mL.
Slant medium: peptone 10g, yeast extract paste 10g, sodium-chlor 10g, agar 16 ~ 20g, water 1000mL, pH nature.
Seed culture medium: peptone 10g, yeast extract paste 10g, sodium-chlor 10g, water 1000mL, pH6.5 ~ 7.2.
Fermention medium: glucose 5g, yeast extract paste 10g, peptone 10g, sodium-chlor 10g, water 1000mL, pH6.0 ~ 7.0.
Shake-flask seed is cultivated: scrape lawn from each bacterial strain inclined-plane respectively and be inoculated in seed culture medium, cultivates 20 ~ 24 hours for 40 DEG C, shaking speed 200 ~ 220rpm.Treat each bacterium liquid OD
600>=1.5, and bottle.And the seed liquor after bottle is inoculated in fermentor tank.
Fermentor cultivation: the in-built 30L fermention medium of 50L fermentor tank, inoculum size 0.1 ~ 0.5%(v/v), cultivate 20 ~ 24 hours for 40 DEG C, fermentor tank rotating speed 100 ~ 200rpm, air flow 1:0.1 ~ 0.3, OD
600>=1.9 stop cultivating.
Microbial inoculum finished product: fermented liquid direct packaging 25kg/ bucket or packing 10kg/ bag after granulating.
The process of embodiment 2 cephalosporin dreg
Ship 40 tons of cephalosporin (CPC) bacterium slags from certain company, water content about 75%, wherein every g dry weight (%) contains about crude protein 42.9, crude fat 2.9, carbohydrate 38.1, robust fibre 3.5, cinder powder 8.7; Wherein, calcium 2.2, phosphorus 1.3; Aminoacids content is in table 1.Cynnematin content is 2414ppm.Add moisture adjuster 20 tons (water content about 30%), fully mix be mixed even, this material moisture about 60%.The thermophilic microorganism microbial inoculum adding in 40L embodiment 1 preparation mix further be again mixed even.Carry out banking up aerobic fermentation at fermenter.1st ~ 3 days, air flow was low, was full dose 700m
3add behind the 1/3 ~ 1/4,4th day of/h that Wind Volume is full dose 1/2 ~ 1, within 8 days, be that full dose is to fermentation ends afterwards.
Every day, 10:00 and 16:00 detected the temperature of material from multiple spot (3 ~ 5), and measured water content in 0,3,5,8,10,13,15,18,20 day with dry weight-loss method, calculate the loss of weight situation of material simultaneously.And press Chinese Pharmacopoeia 2010 editions relevant regulations HPLC methods mensuration CPC content, result is as shown in table 2.
Aminoacids content (dry weight, %) in table 1 cephalosporin bacterium slag
The change (N=4) of each index in table 2 cephalosporin bacterium slag fermenting process
As can be seen from Table 2, releasing heat when decomposing in the growth and breeding process of microorganism under aerobic conditions and utilize a large amount of remaining protein in bacterium slag, sugar, amino acid, inorganic salt and other organism makes heap temperature raise, water content reduces, and heap body weight reduces, and CPC is residual to be reduced.
The process of embodiment 3 terramycin strain slag
Ship 21 tons of terramycin strain slags from certain company, water content about 72%, wherein every g dry weight (%) contains about crude protein 44.7, crude fat 1.1, carbohydrate 36.2, robust fibre 4.3, cinder powder 10.1; Wherein, calcium 5.2, phosphorus 0.3; Aminoacids content is in table 3.Terramycin content 1103ppm in terramycin strain slag.Add moisture adjuster 10 tons (water content about 30%), fully mix be mixed even, this material moisture about 58%.The thermophilic microorganism microbial inoculum adding in 21L embodiment 1 preparation mix further be again mixed even.Be transported in fermenter with forklift and carry out banking up aerobic fermentation.At 1st ~ 4 days of aerobic fermentation, air flow is low was full dose 700m
3add behind the 1/3 ~ 1/4,5th day of/h that Wind Volume is full dose 1/2 ~ 1, within 8 days, be that full dose is to fermentation ends afterwards.
Every day, 10:00 and 16:00 detected the temperature of material from multiple spot (3 ~ 5), and measured water content in 0,3,5,8,10,13,15,18,20 day, calculates the loss of weight situation of material simultaneously.And measuring Penicillin Content by HPLC method, result is as shown in table 4.
Aminoacids content (dry weight, %) in table 3 terramycin strain slag
The change (N=4) of each index in table 4 terramycin strain slag fermenting process
As can be seen from Table 4, releasing heat when decomposing in the process of growth and breeding due to microorganism and utilize the protein in bacterium slag, sugar, inorganic salt and other organism makes heap temperature raise, water content reduces after 10 days fast, the minimizing 20 days time of heap body weight reaches more than 85, and in bacterium slag, oxytetracycline residues declines to a great extent.
The process of embodiment 5 erythromycin bacterium slag
Ship 24 tons of erythromycin bacterium slags from certain company, water content about 68%, wherein every g dry weight (%) contains about crude protein 33.7, crude fat 13.3, carbohydrate 29.7, robust fibre 2.0, cinder powder 13.9; Wherein, calcium 3.4, phosphorus 2.5; Aminoacids content is in table 5.Erythromycin content is about 821ppm.Add moisture adjuster 10 tons (water content 30%), fully mix be mixed even, this material moisture about 57%.The thermophilic microorganism microbial inoculum adding in 24L embodiment 1 preparation mix further be again mixed even.Carry out banking up aerobic fermentation at fermenter.1st ~ 3 days, air flow is low was full dose 700m
3adding Wind Volume behind the 1/3 ~ 1/4,4th day of/h is full dose 700m
3/ h's 1/2 ~ 1,8 days is that full dose is to fermentation ends afterwards.
Every day, 10:00 and 16:00 detected the temperature of material from multiple spot (3 ~ 5), and measured water content in 0,3,5,8,10,13,15,18,20 day with dry weight-loss method, calculate the loss of weight situation of material simultaneously.And press the content that Chinese Pharmacopoeia 2010 editions relevant regulations HPLC methods measure erythromycin in bacterium slag, result is as shown in table 6.
Aminoacids content (dry weight, %) in table 5 erythromycin bacterium slag
The change (N=4) of each index in table 6 erythromycin bacterium slag fermenting process
The process of embodiment 6 lovastatin bacterium slag
Ship 16 tons of lovastatin bacterium slags from certain company, water content about 68%, wherein every g dry weight (%) is about containing crude protein 37.6, crude fat 2.8, carbohydrate 40.7, robust fibre 3.3, cinder powder 8.3; Wherein, calcium 1.2, phosphorus 0.5; Ethyl acetate 3.9; Aminoacids content is in table 7.And be 597ppm containing lovastatin content.Add moisture adjuster 7 tons (water content 30%), fully mix be mixed even, this material moisture about 56%.Mix further after adding in 16L embodiment 1 the thermophilic microorganism microbial inoculum of preparation be mixed even.Aerobic fermentation is carried out at fermenter.1st ~ 3 days, air flow is low was the 700m of full dose
3add behind the 1/3 ~ 1/4,4th day of/h that Wind Volume is full dose 1/2 ~ 1, within 8 days, be that full dose is to fermentation ends afterwards.
Every day, 10:00 and 16:00 detected the temperature of material from multiple spot (3 ~ 5), and measured water content in 0,3,5,8,10,13,15,18,20 day with dry weight-loss method, calculate the loss of weight situation of material simultaneously.And press the content that Chinese Pharmacopoeia 2010 editions relevant regulations HPLC methods measure lovastatin in bacterium slag, result is as shown in table 8.
Aminoacids content (dry weight, %) in table 7 lovastatin bacterium slag
The change (N=4) of each index in table 8 lovastatin bacterium slag fermenting process
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Claims (9)
1., for the treatment of the thermophilic microorganism composite fungus agent of antibiotic bacterium slag, it is characterized in that, in described composite fungus agent effective constituent comprise thermophilic ground bud bacillus that deposit number is CGMCCNo.5641 (
geobacillussp.) UTM01, deposit number be CGMCCNo.5642 thermophilic ground bud bacillus (
geobacillussp.) UTM02, deposit number be CGMCCNo.5643 genus bacillus (
bacillussp.) UTM03, deposit number be CGMCCNo.5927 methylotrophic genus bacillus (
bacillusmethylotrophicus) UTM401, deposit number be CGMCCNo.5928 anaerobic spore-bearing bacilli (
anoxybacillusmongoliensis) UTM501, deposit number be CGMCCNo.5929 thermophilic double oxygen genus bacillus (
anoxybacilluspushchinoensis) UTM601, viable count > 1 × 10 in described thermophilic microorganism composite fungus agent
11cFU/mL.
2. the preparation method of thermophilic microorganism composite fungus agent described in claim 1, is characterized in that, comprises the step of the slant culture of bacterial strain, seed culture and fermentation culture, treats bacterium liquid OD
600>=1.9, merge each bacterium liquid, obtain thermophilic microorganism composite fungus agent.
3. adopt the method for thermophilic microorganism microbial inoculum process antibiotic bacterium slag, it is characterized in that, utilize the thermophilic microorganism composite fungus agent process antibiotic bacterium slag described in claim 1.
4. method according to claim 3, is characterized in that, the moisture content of described antibiotic bacterium slag is 55 ~ 65%, adds microbial inoculum 1L or 1kg in bacterium slag per ton.
5. method according to claim 3, is characterized in that, utilizes described thermophilic microorganism composite fungus agent by aerobic static fermentation technical finesse antibiotic bacterium slag.
6. method according to claim 5, is characterized in that, leavening temperature is more than 90 DEG C, and fermentation period is 15 ~ 20 days.
7. method according to claim 5, is characterized in that, keeps air flow at 1:0.05 ~ 0.2v/v in fermenting process.
8. method according to claim 3, is characterized in that, described antibiotic bacterium slag includes but not limited to cephalosporin dreg, terramycin strain slag, erythromycin bacterium slag and lovastatin bacterium slag.
9. method according to claim 3, is characterized in that, also comprises the step that the bacterium slag after to microbial inoculum process burns.
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| CN106694521B (en) * | 2017-01-16 | 2018-09-11 | 河北科技大学 | One metal ion species strengthen method of the thermophilic lysis bacterium for antibiotic bacterium dregs minimizing |
| CN107413821A (en) * | 2017-08-10 | 2017-12-01 | 河北科技大学 | A kind of method that the thermophilic lysis bacterium of immobilization is used for antibiotic bacterium dregs minimizing |
| CN108033817B (en) * | 2017-11-27 | 2019-11-26 | 福建农林大学 | A kind of method of antibiotic and resistant gene in quick abatement organic solid waste |
| CN109337837B (en) * | 2018-11-06 | 2021-07-23 | 河北科技大学 | Bacillus thermophilus DF7 strain and application thereof |
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